Oxidative Stress And MAPKs Signaling Pathway In Ovarian Granulosa Cells Of Infertity Patients With Different TCM Constitution

ObjectiveTo explore the relationship between oxidative stress, MAPKs signaling pathway and different TCM Constitution types of ovary granulosa cells in infertility patients.MethodsA cross-sectional survey research method is used. And then doctor coll ect the follicular fluid, social information,clinical symptoms and the rel ated clinical information of the 22 patients who have infertility and und ergoing the IVF or ICSI.Detect MDA and SOD in follicular fluid with reage nt kit. Western blotting was adopted to detect the differences of the exp ressional level of MAPK signaling pathway factor P-SAPK/Juk, P-P38 and P-E RK1/2 protein between different age and constitution groups. Patients wer e divided into deficiency,excess and gentleness constitution, the differe nces in the results between the three groups were compared. By age of 30 y ears old, patients were divided into two groups, the differences in the r esults between the two groups were compared. A database was established b y using Excel or SPSS 19.0 statistical package for preliminary exploratio n on the relationship between oxidative stress,MAPK signaling pathway and TCM constitution.Results1.The TCM constitutions of infertility were devided into three typesa nd they are distributed as follows:most of deficiency constitution in 12 cases(56.5%), followed excess constitution in 3 cases(30.4%), gentleness c onstitution in 3 cases(13%).2. The MDA content in follicular fluid was compared between the three constitution groups, the results showed that there was no difference betw een groups (P>0.05). The SOD content in follicular fluid was compared bet ween the three constitution groups, the results showed that there was no difference between groups (P>0.05).3.The MDA content in follicular fluid was compared between the two ag e groups, the results showed that there was no difference between groups (P>0.05). The SOD content in follicular fluid was compared between the tw o age groups, the results showed that there was no difference between gro ups(P>0.05).4.11 patients were picked out of the 22 patients, whose ovulation induction scheme were long scheme, infertility factor were male factor or gamete transport barriers. Since only one patient has gentleness constitution, this patient was picked out. P-SAPK/Juk in granulosa cells were compared in deficiency and excess constitution.. There was no significant difference between the two groups of P-SAPK/Juk (P>0.05). Due to the limitation of the sample protein, only 14 patients were detected by P-P38. There was no significant difference in the absorbance of P-P38 between the three constitution groups (P>0.05). Due to the limitation of the sample protein, only 9 patients were detected by P-ERK1/2. There was no significant difference in the absorbance of P-ERK1/2 between the three constitution groups (P>0.05).5. The P-SAPK/Jnk content in granulosa cells were compared between the two age groups. The results showed that there was significant difference between groups at the level of P<0.05(P=0.002).There was no significant difference in the absorbance of P-P38 between the two age groups (P>0.05).There was no significant difference in the absorbance of P-ERK1/2 between the two age groups (P>0.05).Conclusion1. There was no significant difference in MDA and SOD levels in follicular fluid between different constitution groups and age groups.2. MAPK/Jnk signaling pathway may be involved in the process of oxidative stress in granulosa cells. In our study we temporarily find no significant difference of MAPK/Jnk expressed in different constitution patients.3. In our study we temporarily find no significant difference of P-P38 and P-ERK1/2 expressed in different constitution and age patients. Further study is needed to explore the role of P-P38 and P-ERK1/2 in oxidative stress pathways in granulosa cells.